Immunomodulatory Peptides

ABSTRACT

This disclosure provides peptides which can therefore be used for various therapeutic purposes, such as inhibiting the progression of a hyperproliferative disorder, including cancer; treating infectious diseases; enhancing a response to vaccination; treating sepsis; and promoting hair re-pigmentation or lightening of pigmented skin lesions.

This application incorporates by reference the contents of a 1.52 kb text filed created on Oct. 7, 2020 and named “00047900276sequencelisting.txt,” which is the sequence listing for this application.

Each scientific reference, patent, and published patent application cited in this disclosure is incorporated herein by reference in its entirety.

TECHNICAL FIELD

This disclosure relates generally to immunomodulatory peptides.

BACKGROUND

Programmed cell death-1 (PD1) and its ligands, PD-L1 and PD-L2, are widely expressed and exert a number of immunoregulatory roles in T cell activation, including attenuation of immunity against tumor cells and infectious agents. PD1 is therefore an attractive target for a variety of therapeutic applications. There is a continuing need for useful modulators of immune checkpoint pathways.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a graph showing the ability of various peptides to inhibit the activity of PD1 to recruit SHP1 using PATHHUNTER® checkpoint signaling assays (DiscoverX).

FIG. 2 is a graph showing how peptides increase the number of IFN_(γ)-positive CD8 T cells measured by IFN_(γ)-ELISPOT when combined with a vaccine antigen (AdPyCS).

FIG. 3A, graph comparing MC38 tumor volume in mice, normalized to day 0 values for each animal. Cohorts of mice received intra-tumoral (IT) injections of 50 μg FoLD01 peptide or intraperitoneal injections of antibodies against CTLA4 and/or PD1. The median value for control cohort (DMSO/PBS injected intra-tumorally) is shown as a heavier line.

FIG. 3B, graph comparing normalized tumor growth kinetics for the indicated groups. Error bars represent cohort mean and 95% confidence intervals.

FIG. 3C, graph comparing normalized tumor growth at day 16, p values determined using non-parametric Mann-Whitney t-test.

FIG. 4A, graph comparing Pan02 tumor volume in mice, normalized to day 0 values for each animal. Cohorts of mice received intra-tumoral injections of 50 μg FoLD04 or intraperitoneal injections of antibodies against CTLA4 and/or PD1. The median value for control cohort (DMSO/PBS injected intra-tumorally) is shown as a heavier line.

FIG. 4B, graph comparing normalized tumor growth kinetics for the indicated groups. Error bars represent cohort mean and 95% confidence intervals.

FIG. 4C, graph comparing normalized tumor growth at day 21, p values determined using non-parametric Mann-Whitney t-test.

FIG. 4D, graph comparing normalized tumor growth at day 27, p values determined using non-parametric Mann-Whitney t-test.

DETAILED DESCRIPTION

This disclosure provides peptides that antagonize the activity of the checkpoint receptor “programmed death 1” (PD-1). The amino acid sequences of these peptides are shown below.

peptide amino acid sequence SEQ ID NO: FoLD01 STNQVSALRVNILFPLSQ 1 FoLD02 STNQVSALKVNILFPLSQ 2 FoLD03 STGQVSTLRVNITAPLSQ 3 FoLD04 QVSALRVNILF 4

In some embodiments, the disclosed peptides consist of the amino acid sequence of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4. In some embodiments, the disclosed peptides comprise the amino acid sequence of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4. In some embodiments, the disclosed peptides consist essentially of the amino acid sequence of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4.

In some embodiments, a disclosed peptide is modified using chemical or recombinant methods to enhance its stability or other pharmacokinetic properties. See, e.g., US 2017/0020956. Modifications include, but are not limited to, replacement of one or more L-amino acid with its corresponding D-form, acetylation on a C- and/or N-terminal residue, amidation on a C- and/or N-terminal residue, cyclization, esterification, glycosylation, acylation, attachment of myristic or palmitic acid, addition of an N-terminal glycine, addition of lipophilic moieties such as long fatty acid chains, and PEGylation.

Peptides can be made by any method known in the art, including synthetic methods, recombinant methods, or both. Synthetic methods include solid-phase and solution methods, and may include the use of protective groups. See, e.g., Bodanszky et al. (1976), McOmie (1973), Merrifield (1963), Neurath et al. (1976), Stuart & Young (1984).

Recombinant production of unmodified peptides can be carried out using any nucleotide sequence(s) encoding the peptides in any suitable expression system. Nucleic acid molecules encoding one or more of the disclosed peptides can be incorporated into an expression cassette that includes control elements operably linked to the coding sequences. Control elements include, but are not limited to, initiators, promoters (including inducible, repressible, and constitutive promoters), enhancers, and polyadenylation signals. Signal sequences can be included. The expression cassette can be provided in a vector that can be introduced into an appropriate host cell for production of the peptide(s). Methods of constructing expression cassettes and expression vectors are well known. Expression vectors can include one or more expression cassettes encoding one or more peptides comprising, consisting essentially of, or consisting of SEQ ID NO:1, 2, 3, or 4.

In some embodiments, a disclosed peptide, or modified version thereof, is conjugated to a moiety, such as albumin or transthyretin, to enhance the plasma half-life of the peptide. Methods of preparing such conjugates are well known in the art (e.g., Penchala et al., 2015; Kontermann, 2016; Zorzi et al., 2017).

In some embodiments, a disclosed peptide, or modified version thereof, is conjugated to a partner molecule, such as a peptide or protein such as an antibody intended to increase the half-life of the peptide or modified peptide in vivo and/or to provide specific delivery to a target tissue or cell. Conjugation may be direct or can be via a linker. In some of these embodiments, a peptide or a modified version thereof can be altered to substitute one or more amino acids with amino acids used to attach partner molecules, such as lysine, or by N-terminal extension of the peptide with, e.g., 1, 2, 3, or 4 glycine spacer molecules.

This disclosure also provides CAR-T cells that express one or more of the disclosed peptides. Methods of preparing CAR-T cells are disclosed, for example, in U.S. Pat. Nos. 9,328,156; 9,845,362; and 9,101,584.

This disclosure also provides oncolytic viruses containing a nucleic acid molecule encoding one or more of the disclosed peptides. See US 2017/0157188; Lawler et al., 2017; US 2015/0250837. Oncolytic viruses include, but are not limited to, reovirus, Seneca Valley virus, vesicular stomatitis virus, Newcastle disease virus, herpes simplex virus, morbillivirus virus, retrovirus, influenza virus, Sindbis virus, poxvirus, and adenovirus.

Examples of oncolytic reovirus include REOLYSIN® (pelareorep) and reoviruses disclosed in US 2017/0049829.

Examples of oncolytic Seneca Valley virus include NTX-101 (Rudin et al., 2011).

Examples of oncolytic vesicular stomatitis virus are disclosed in Stojdl et al., 2000; and Stojdl et al., 2003.

Examples of oncolytic Newcastle disease virus include 73-T PV701 and HDV-HUJ strains (see also Phuangsab et al., 2001; Lorence et al., 2007; and Freeman et al., 2006).

Examples of oncolytic herpes simplex virus include NV1020 (Geevarghese et al., 2010) and T-VEC (Andtbacka et al., 2013).

Examples of oncolytic morbillivirus virus include oncolytic measles viruses such as MV-Edm (McDonald et al., 2006) and HMWMAA (Kaufmann et al., 2013).

Examples of oncolytic retrovirus are disclosed in Lu et al., 2012.

Examples of oncolytic influenza virus are disclosed, for example, in US 2018/0057594.

Examples of oncolytic Sindbis virus are disclosed, for example, in Lundstrom, 2017.

Examples of oncolytic poxvirus are disclosed, for example, in Chan & McFadden, 2014.

Examples of oncolytic adenovirus include ONYX-015 (Khuri et al., 2000) and H101 or Oncorine (Liang, 2018).

Therapeutic Uses

The disclosed peptides have a number of therapeutic applications, including treating hyperproliferative disorders, including cancer, treating infectious diseases, enhancing a response to vaccination, treating sepsis, promoting hair re-pigmentation, and promoting lightening of a pigmented skin lesion. “Treat,” as used herein, includes reducing or inhibiting the progression of one or more symptoms of the condition for which a peptide or modified version thereof is administered.

“Administer” as used herein includes direct administration of a disclosed peptide or modified version thereof as well as indirect administration.

In some embodiments, one or more of the disclosed peptides are directly administered. In some of these embodiments, a peptide carrier system is used. A number of peptide carrier systems are known in the art, including microparticles, polymeric nanoparticles, liposomes, solid lipid nanoparticles, hydrophilic mucoadhesive polymers, thiolated polymers, polymer matrices, nanoemulsions, and hydrogels. See Patel et al. (2014), Bruno et al. (2013), Feridooni et al. (2016). Any suitable system can be used.

In some embodiments, an engineered T cell that expresses and secretes one or more disclosed peptides can be used to deliver PD1 inhibition at the site of engagement of the T cell receptor with an antigen. The T cell-based therapy can be, for example, a CAR-T cell that expresses one or more of the disclosed peptides. Either inducible or constitutive expression can be used.

In some embodiments, an oncolytic virus can be used to deliver one or more of the disclosed peptides. Either inducible or constitutive expression can be used.

In other embodiments one or more of the disclosed peptides are delivered using one or more nucleic acids encoding the peptide(s) (e.g., DNA, cDNA, PNA, RNA or a combination thereof); see, e.g., US 2017/0165335. Nucleic acids encoding one or more peptides can be delivered using a variety of delivery systems known in the art. Nucleic acid delivery systems include, but are not limited to, gene-gun; cationic lipids and cationic polymers; encapsulation in liposomes, microparticles, or microcapsules; electroporation; virus-based, and bacterial-based delivery systems. Virus-based systems include, but are not limited to, modified viruses such as adenovirus, adeno-associated virus, herpes virus, retroviruses, vaccinia virus, or hybrid viruses containing elements of one or more viruses. US 2002/0111323 describes use of “naked DNA,” i.e., a “non-infectious, non-immunogenic, non-integrating DNA sequence,” free from “transfection-facilitating proteins, viral particles, liposomal formulations, charged lipids and calcium phosphate precipitating agents,” to administer a peptide. Bacterial-based delivery systems are disclosed, e.g., in Van Dessel et al. (2015) and Yang et al. (2007).

In some embodiments, a peptide is administered via an RNA molecule encoding the peptide. In some embodiments, the RNA molecule is encapsulated in a nanoparticle. In some embodiments, the nanoparticle comprises a cationic polymer (e.g., poly-L-lysine, polyamidoamine, polyethyleneimine, chitosan, poly(β-amino esters). In some embodiments, the nanoparticle comprises a cationic lipid or an ionizable lipid. In some embodiments, the RNA molecule is conjugated to a bioactive ligand (e.g., N-acetylgalactosamine (GalNAc), cholesterol, vitamin E, antibodies, cell-penetrating peptides). See, e.g., Akinc et al. (2008), Akinc et al. (2009), Anderson et al. (2003), Behr (1997), Boussif et al. (1995), Chen et al. (2012), Dahlman et al. (2014), Desigaux et al. (2007), Dong et al. (2014), Dosta et al. (2015), Fenton et al. (2016), Guo et al. (2012), Howard et al. (2006), Kaczmarek et al. (2016), Kanasty et al. (2013), Kauffman et al. (2015), Kozielski et al. (2013), Leus et al. (2014), Lorenz et al. (2004), Love et al. (2010), Lynn & Langer (2000), Moschos et al. (2007), Nair et al. (2014), Nishina et al. (2008), Pack et al. (2005), Rehman et al. (2013), Schroeder et al. (2010), Tsutsumi et al. (2007), Tzeng et al. (2012), Won et al. (2009), Xia et al. (2009), Yu et al. (2016).

-   In some embodiments, an RNA molecule can be modified to reduce its     chances of degradation or recognition by the immune system. The     ribose sugar, the phosphate linkage, and/or individual bases can be     modified. See, e.g., Behlke (2008), Bramsen (2009), Chiu (2003),     Judge & MacLachlan (2008), Kauffman (2016), Li (2016), Morrissey     (2005), Prakash (2005), Pratt & MacRae (2009), Sahin (2014),     Soutschek (2004), Wittrup & Lieberman (2015). In some embodiments,     the modification is one or more of a ribo-difluorotoluyl nucleotide,     a 4′-thio modified RNA, a boranophosphate linkage, a     phosphorothioate linkage, a 2′-O-methyl (2′-OMe) sugar substitution,     a 2′-fluoro (2′-F), a 2′-O-methoxyethyl (2′-MOE) sugar substitution,     a locked nucleic acid (LNA), and an L-RNA.

In some embodiments, administration is carried out in conjunction with one or more other therapies. “In conjunction with” includes administration together with, before, or after administration of the one or more other therapies.

Pharmaceutical Compositions, Routes of Administration, and Devices

One or more peptides, nucleic acid molecules, CAR-T cells, and/or oncolytic viruses, as discussed above, are typically administered in a pharmaceutical composition comprising a pharmaceutically acceptable vehicle. The “pharmaceutically acceptable vehicle” may comprise one or more substances which do not affect the biological activity of the peptides or modified versions thereof and, when administered to a patient, does not cause an adverse reaction. Pharmaceutical compositions may be liquid or may be lyophilized. Lyophilized compositions may be provided in a kit with a suitable liquid, typically water for injection (WFI) for use in reconstituting the composition. Other suitable forms of pharmaceutical compositions include suspensions, emulsions, and tablets.

Pharmaceutical compositions can be administered by any suitable route, including, but not limited to, intravenous, intramuscular, intradermal, intraperitoneal, subcutaneous, epidural, intratumoral, transdermal (e.g., US 2017/0281672), mucosal (e.g., intranasal or oral), pulmonary, and topical (e.g., US 2017/0274010) routes. See, e.g., US 2017/0101474.

Administration can be systemic or local. In addition to local infusions and injections, implants can be used to achieve a local administration. Examples of suitable materials include, but are not limited to, sialastic membranes, polymers, fibrous matrices, and collagen matrices.

Topical administration can be by way of a cream, ointment, lotion, transdermal patch (such as a microneedle patch), or other suitable forms well known in the art.

Administration can also be by controlled release, for example, using a microneedle patch, pump and/or suitable polymeric materials. Examples of suitable materials include, but are not limited to, poly(-hydroxy ethyl methacrylate), poly(methyl methacrylate), poly(acrylic acid), poly(ethylene-co-vinyl acetate), poly(methacrylic acid), polyglycolides (PLG), polyanhydrides, poly(N-vinyl pyrrolidone), poly(vinyl alcohol), polyacrylamide, poly(ethylene glycol), polylactides (PLA), poly(lactide-co-glycolides) (PLGA), and polyorthoesters.

Devices comprising any of the peptides, nucleic acid molecules, CAR-T cells, and/or oncolytic viruses described above include, but are not limited to, syringes, pumps, transdermal patches, spray devices, vaginal rings, and pessaries.

Treatment of Hyperproliferative Disorders, Including Cancer

In some embodiments, one or more of the peptides, nucleic acid molecules, CAR-T cells, and/or oncolytic viruses described above are administered to a patient to inhibit the progression of a hyperproliferative disorder, including cancer. Such inhibition may include, for example, reducing proliferation of neoplastic or pre-neoplastic cells; destroying neoplastic or pre-neoplastic cells; and inhibiting metastasis or decreasing the size of a tumor.

Examples of cancers include, but are not limited to, melanoma (including cutaneous or intraocular malignant melanoma), renal cancer, prostate cancer, breast cancer, colon cancer, lung cancer, bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, Hodgkin's Disease, non-Hodgkin's lymphoma, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, chronic or acute leukemias including acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, lymphocytic lymphoma, cancer of the bladder, cancer of the kidney or ureter, carcinoma of the renal pelvis, neoplasm of the central nervous system (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid cancer, squamous cell cancer, and T-cell lymphoma.

Combination Cancer Therapies

In some embodiments, one or more of the peptides, nucleic acid molecules, CAR-T cells, and/or oncolytic viruses described above are administered in conjunction with one or more other cancer therapies or immunotherapies, such as those described below.

In some embodiments, the second therapy comprises a second agent that reduces or blocks the activity of PD1 (e.g., nivolumab, pembrolizumab, durvalumab) or CTLA-4 (e.g., ipilimumab, tremelimumab).

In some embodiments, the second therapy comprises an agent that reduces or blocks the activity of PD-L1 (e.g., atezolizumab).

In some embodiments, the second therapy comprises an agent that reduces or blocks the activity of other inhibitory checkpoint molecules and/or molecules that suppress the immune system. These molecules include, but are not limited to:

-   -   1. Lymphocyte-activation gene-3 (LAG-3; see He et al., 2016;         Triebel et al., 1990);     -   2. V-domain Immunoglobulin Suppressor of T cell Activation         (VISTA, also known as c10orf54, PD1H, DD1α, Gi24, Dies1, and         SISP1; see US 2017/0334990, US 2017/0112929, Gao et al., 2017,         Wang et al., 2011; Liu et al., 2015);     -   3. T-cell Immunoglobulin domain and Mucin domain 3 (TIM-3; see         US 2017/0198041, US 2017/0029485, US 2014/0348842, Sakuishi et         al., 2010);     -   4. killer immunoglobulin-like receptors (KIRs; see US         2015/0290316);     -   5. agents that inhibit indoleamine (2,3)-dioxygenase (IDO; see         Mellemgaard et al., 2017);     -   6. B and T Lymphocyte Attenuator (BTLA; see US 2016/09222114);         and     -   7. A2A adenosine receptor (A2AR; see Beavis et al., 2015; US         2013/0267515; US 2017/0166878; Leone et al., 2015;         Mediavilla-Varela et al., 2017; Young et al., 2016).

Agents that reduce or block the activity of LAG-3 include, but are not limited to, BMS-986016, IMP321, and GSK2831781 (He et al., 2016).

Agents that reduce or block the activity of VISTA include, but are not limited to, small molecules, such as CA-170, and antibodies (e.g., Le Mercier et al., 2014).

Agents that reduce or block the activity of TIM-3 include, but are not limited to, antibodies such as MBG453 and TSR-022; see Dempke et al., 2017.

Agents that reduce or block the activity of KIRs include, but are not limited to, monoclonal antibodies such as IPH2101 and Lirilumab (BMS-986015, formerly IPH2102); see Benson & Caligiuri, 2014.

Agents that reduce or block the activity of IDO include, but are not limited to, epacadostat and agents disclosed in US 2017/0037125.

Agents that reduce or block the activity of BTLA include, but are not limited to, peptides (e.g., Spodzieja et al., 2017).

Agents that reduce or block the activity of A2AR include, but are not limited to, small molecules such as CPI-444 and vipadenant.

In some embodiments, the second therapy comprises a cytokine (e.g., interleukin 7).

In some embodiments, the second therapy comprises an agonist of a stimulatory checkpoint molecule. These molecules include, but are not limited to:

-   -   1. CD40;     -   2. OX40;     -   3. glucocorticoid-induced tumor necrosis factor-related protein         (GITR); and     -   4. Inducible T-cell COStimulator (ICOS).

Agonists of CD40 include, but are not limited to, CD40 agonist monoclonal antibodies such as cp-870,893, ChiLob7/4, dacetuzumab, and lucatumumab. See, e.g., Vonderheide et al., 2007; Khubchandani et al., 2009; Johnson et al., 2010; Bensinger et al., 2012; Vonderheide and Glennie, 2013; Johnson et al., 2015.

Agonists of OX40 include, but are not limited to, OX40 agonist antibodies such as MOXR0916, MED16469, MED10562, PF-045618600, GSK3174998, and INCCAGN01949, and OX40L-Fc fusion proteins, such as MEDI6383. See, e.g., Huseni et al., 2014; Linch et al., 2015; Messenheimer et al., 2017. See also Shrimali et al., 2017.

Agonists of GITR include, but are not limited to, MEDI1873. See, e.g., Schaer et al., 2012; Tigue et al., 2017.

Agonists of ICOS include, but are not limited to, ICOS agonist antibodies JTX-2011 and GSK3359609. See, e.g., Harvey et al., 2015; Michaelson et al., 2016.

In other embodiments, the second therapy comprises a 4-1BB agonist (Shindo et al., 2015), such as urelumab; a 4-1BB antagonist (see US 2017/0174773); an inhibitor of anaplastic lymphoma kinase (ALK; Wang et al., 2014; US 2017/0274074), such as crizotinib, ceritinib, alectinib, PF-06463922, NVP-TAE684, AP26113, TSR-011, X-396, CEP-37440, RXDX-101; an inhibitor of histone deacetylase (HDAC; see US 2017/0327582); a VEGFR inhibitor, such as axitinib, sunitinib, sorafenib, tivozanib, bevacizumab; and/or an anti-CD27 antibody, such as varlilumab.

In some embodiments, the second therapy comprises a cancer vaccine (e.g., Duraiswamy et al., 2013). A “cancer vaccine” is an immunogenic composition intended to elicit an immune response against a particular antigen in the individual to which the cancer vaccine is administered. A cancer vaccine typically contains a tumor antigen which is able to induce or stimulate an immune response against the tumor antigen. A “tumor antigen” is an antigen that is present on the surface of a target tumor. A tumor antigen may be a molecule which is not expressed by a non-tumor cell or may be, for example, an altered version of a molecule expressed by a non-tumor cell (e.g., a protein that is misfolded, truncated, or otherwise mutated).

In some embodiments, the second therapy comprises a chimeric antigen receptor (CAR) T cell therapy. See, e.g., John et al., 2013; Chong et al., 2016.

In some embodiments, one or more of the peptides, nucleic acid molecules, CAR-T cells, and/or oncolytic viruses described above are administered in conjunction with a CAR-T cell cancer therapy to increase the efficacy of the CAR-T cell cancer therapy.

In some embodiments, one or more of the peptides, nucleic acid molecules, CAR-T cells, and/or oncolytic viruses described above are administered in conjunction with an oncolytic virus as disclosed, for example, in US 2017/0143780. Non-limiting examples of oncolytic viruses are described above.

Additional Therapeutic Uses

In some embodiments, one or more of the peptides, nucleic acid molecules, CAR-T cells, and/or oncolytic viruses described above are administered to a patient to treat infectious diseases, including chronic infections, caused, e.g., by viruses, fungi, bacteria, and protozoa, and helminths.

Examples of viral agents include human immunodeficiency virus (HIV), Epstein Barr Virus (EBV), Herpes simplex (HSV, including HSV1 and HSV2), Human Papillomavirus (HPV), Varicella zoster (VSV) Cytomegalovirus (CMV), and hepatitis A, B, and C viruses.

Examples of fungal agents include Aspergillus, Candida, Coccidioides, Cryptococcus, and Histoplasma capsulatum.

Examples of bacterial agents include Streptococcal bacteria (e.g., pyogenes, agalactiae, pneumoniae), Chlamydia pneumoniae, Listeria monocytogenes, and Mycobacterium tuberculosis.

Examples of protozoa include Sarcodina (e.g., Entamoeba), Mastigophora (e.g., Giardia), Ciliophora (e.g., Balantidium), and Sporozoa (e.g., Plasmodium falciparum, Cryptosporidium).

Examples of helminths include Platyhelminths (e.g., trematodes, cestodes), Acanthocephalins, and Nematodes.

In some embodiments, one or more of the peptides, nucleic acid molecules, CAR-T cells, and/or oncolytic viruses described above are administered as a vaccine adjuvant, to enhance a response to vaccination (e.g., by increasing effector T cells and/or reducing T cell exhaustion). The vaccine can be, for example, an RNA vaccine (e.g., US 2016/0130345, US 2017/0182150), a DNA vaccine, a recombinant vector, a protein vaccine, or a peptide vaccine. Such vaccines can be delivered, for example, using virus-like particles, as is well known in the art.

In some embodiments, one or more of the peptides, nucleic acid molecules, CAR-T cells, and/or oncolytic viruses described above are administered to treat sepsis.

In some embodiments, one or more of the peptides, nucleic acid molecules, CAR-T cells, and/or oncolytic viruses described above are administered to promote hair color re-pigmentation. In some embodiments, one or more of the peptides, nucleic acid molecules, CAR-T cells, and/or oncolytic viruses described herein are administered to promote lightening of pigmented skin lesions.

Example 1. PATHHUNTER® Checkpoint Signaling Assays

Peptides were tested for their ability to inhibit the binding of PDL1 to PD1 using PATHHUNTER® checkpoint signaling assays (DiscoverX).

Jurkat cells expressing PD1 and SHP1 proteins, each fused to a fragment of enzyme fragment complementation (EFC) system, were co-incubated with PDL1-presenting U2OS cells. This results in PD1 activation and SHP1 recruitment to the PD1 receptors, bringing together the two EFC fragments and generating a light signal. Inhibitory peptides or antibodies added to the culture would reduce the light signal.

Cells in co-culture were incubated at room temperature (RT) for 2 h (PD1 assay). The assay signal was generated using the PATHHUNTER® Bioassay Detection kit. FoLD01 (SEQ ID NO:1), LD10 (SEQ ID NO:5), and LD12 (SEQ ID NO:6) peptides were tested in duplicate at two concentrations, 20 μM and 100 μM. Peptides were dissolved in water or DMSO and diluted in assay buffer.

Microplates were read following signal generation using a PerkinElmer ENVISION™ instrument for chemiluminescent signal detection. The percentage of inhibition efficacy was calculated using the following formula, in which “RLU” means relative light units:

$100 \times \left\lbrack {1 - \left\lbrack \frac{\begin{matrix} {\left( {{mean}\mspace{14mu}{RLU}\mspace{14mu}{of}\mspace{14mu}{test}\mspace{14mu}{sample}} \right) -} \\ \left( {{mean}\mspace{14mu}{RLU}\mspace{14mu}{of}\mspace{14mu}{vehicle}\mspace{14mu}{control}} \right) \end{matrix}}{\begin{matrix} {\left( {{mean}\mspace{14mu}{RLU}\mspace{14mu}{of}\mspace{14mu}{EC}_{80}\mspace{14mu}{control}} \right) -} \\ \left( {{mean}\mspace{14mu}{RLU}\mspace{14mu}{of}\mspace{14mu}{vehicle}\mspace{14mu}{control}} \right) \end{matrix}} \right\rbrack} \right\rbrack$

The results demonstrate that the FoLD01 peptide inhibited PD1-PDL1 interaction to a greater extent than peptides LD10 and LD12 (FIG. 1).

Example 2. Evaluation of Peptides in an AdPyCS Mouse Model

Groups of five mice were immunized intramuscularly with a recombinant replication defective adenovirus expressing the Plasmodium yoelli circumsporozoite protein (AdPyCSP) and then treated subcutaneously with peptides LD10da (SEQ ID NO:6, in which the first amino acid is a D-amino acid, 1 μg), FoLD01 (1 μg, 10 μg), or an anti-PD1 monoclonal antibody (10 μg). Ten days post-injection, mice were euthanized and individual spleens removed. The number of IFNγ-secreting antigen-specific CD8+ T cells were determined using an ELISPOT assay (FIG. 2).

The results showed that peptide FoLD01 enhances the number of antigen-specific CD8+ T cells relative to stimulation with antigen only (one-way ANOVA, statistical differences are relative to AdPyCS-only stimulation).

Example 3. Assessment of Peptides in an MC38 Tumor Model

Mice were injected subcutaneously with 1×10⁶ MC38 (colon carcinoma) cells and tumor volumes were measured on day 2, 5, 9, 12, and 16 (end of study) and normalized to day 0 values for each animal. When the mean tumor size reached approximately 80-120 mm³, mice were randomized and treatment was started. Peptides were tested in an MC38 tumor model. FoLD01 peptide was administered intra-tumorally 2× a week (50 μg dose). Monoclonal antibodies were administered intraperitoneally 2× a week (5 mg/kg (anti-CTLA4) and 10 mg/kg (anti-PD1) dose). Statistical test used: Unpaired, non-parametric, two-tailed Mann-Whitney test. [85] Control used: intra-tumor injection of DMSO diluted in PBS (for FoLD01 injected intra-tumorally). The results are shown in FIG. 3 (A-C). In general, administration of anti-CTLA4 mAb and anti-CTLA4+anti-PD1 mAb resulted in significant decreases in tumor volume. Administration of anti-PD1 mAb resulted in a limited decrease in tumor volume, which may have been attributed to the 1×10⁶ cells MC38 cells injected.

Intra-tumoral injection of FoLD01 peptide significantly decreased tumor volume relative to the intra-tumoral DMSO/PBS control using the non-parametric Mann-Whitney t-test. The P value was 0.0499.

Example 4. Assessment of Peptides in a Pan02 Tumor Model

Mice were injected subcutaneously with 3×10⁶ Pan02 (pancreatic adenocarcinoma) cells in the right rear flank. Tumor volumes were measured at least twice weekly in two dimensions using a caliper. When the mean tumor size reached approximately 80-120 mm³, mice were randomized and treatment was started. FoLD04 was administered intra-tumorally 2× a week (50 μg dose). Monoclonal antibodies were administered intraperitoneally 2× a week (5 mg/kg (anti-CTLA4) and 10 mg/kg (anti-PD1) dose).

Tumor volumes were measured on day 4, 7, 11, 14, 18, 21, 25 and 27 (end of study) and normalized to day 0 values for each animal. Statistical test used: Unpaired, non-parametric, two-tailed Mann-Whitney test.

Control used: intra-tumor injection of DMSO diluted in PBS (for FoLD04 injected intra-tumorally). The results are shown in FIG. 4.

Summary and Interpretation of Pan02 Model Results

Treatment with anti-CTLA4 mAb and anti-CTLA4+anti-PD1 mAb resulted in significant decreases in tumor volume regardless of time point assessed and statistical test used. The anti-PD1 mAb resulted in sporadic significant decreases in tumor volume depending on time point assessed and statistical test used.

When administered intra-tumorally, the FoLD04 peptide demonstrated significantly decreased tumor volume relative to DMSO/PBS IT control using the non-parametric Mann-Whitney t-test.

Based on the time points assessed and the statistical tests used, subcutaneous administration of the peptides trended toward control of tumor volume but did not approach statistical significance.

REFERENCES

-   Adams et al., “Big opportunities for small molecules in     immuno-oncology,” Nature Reviews Drug Discovery Advance Online     Publication, Jul. 31, 2016, 20 pages -   Akinc et al., “A combinatorial library of lipid-like materials for     delivery of RNAi therapeutics,” Nat. Biotechnol. 26, 561-69, 2008 -   Akinc et al., “Development of lipidoid-siRNA formulations for     systemic delivery to the liver,” Mol. Ther. 17, 872-79, 2009 -   Alsaab et al., “PD1 and PD-L1 Checkpoint Signaling Inhibition for     Cancer Immunotherapy: Mechanism, Combinations, and Clinical     Outcome,” Front. Pharmacol. 8, 561, 2017 -   Anderson et al., “semi-automated synthesis and screening of a large     library of degradable cationic polymers for gene delivery,” Angew.     Chemi Int. Ed. 42, 3153-58, 2003 -   Andtbacka et al., “OPTiM: A randomized phase III trial of talimogene     laherparepvec (T-VEC) versus subcutaneous (SC)     granulocyte-macrophage colony-stimulating factor (GM-CSF) for the     treatment (tx) of unresected stage IIIB/C and IV melanoma,” J. Clin.     Oncol. 31, abstract number LBA9008, 2013 -   Beavis et al., “Adenosine Receptor 2A Blockade Increases the     Efficacy of Anti-PD1 through Enhanced Antitumor T-cell Responses,”     Cancer Immunol. Res. 3, 506-17, 2015 -   Behlke, “Chemical modification of siRNAs for in vivo use,”     Oligonucleotides. 2008; 18:305-19. -   Behr, “The proton sponge: a trick to enter cells the viruses did not     exploit,” Int. J. Chem. 2, 34-36, 1997 -   Bensinger et al., “A phase 1 study of lucatumumab, a fully human     anti-CD40 antagonist monoclonal antibody administered intravenously     to patients with relapsed or refractory multiple myeloma,” Br J     Haematol. 159, 58-66, 2012. -   Benson & Caligiuri, “Killer Immunoglobulin-like Receptors and Tumor     Immunity,” Cancer Immunol Res 2014; 2:99-104 -   Bodanszky et al., Peptide Synthesis, John Wiley and Sons, 2d ed.     (1976) -   Boussif et al., “A versatile vector for gene and oligonucleotide     transfer into cells in culture and in vivo: polyethylenimine,” Proc.     Nat'l. Acad. Sci. (USA) 92, 7297-301, 1995 -   Bramsen et al., “A large-scale chemical modification screen     identifies design rules to generate siRNAs with high activity, high     stability and low toxicity,” Nucleic Acids Res. 2009; 37:2867-81 -   Bruno et al., “Basics and recent advances in peptide and protein     drug delivery,” Ther. Deliv. 4, 1443-67, 2013 -   Bu et al., “Learning from PD1 Resistance: New Combination     Strategies,” Trends Mol. Med. 22, 448-51, 2016 -   Burnett & Rossi, “RNA-based Therapeutics—Current Progress and Future     Prospects,” Chem Biol. 19, 60-71, 2012 -   Cao, “Advances in Delivering Protein and Peptide Therapeutics,”     Pharmaceutical Technology 40, 22-24, Nov. 2, 2016 -   Chan & McFadden, “Oncolytic Poxviruses,” Ann. Rev. Virol. 1, 119-41,     2014 -   Chen et al., “Rapid discovery of potent siRNA-containing lipid     nanoparticles enabled by controlled microfluidic formulation,” J.     Am. Chem. Soc. 134, 6948-51, 2012 -   Cherkassky et al., “Human CAR T cells with cell-intrinsic PD1     checkpoint blockade resist tumor-mediated inhibition,” J. Clin.     Invest. 126, 3130-44, 2016 -   Chiu et al., “siRNA function in RNAi: a chemical modification     analysis,” RNA 2003; 9:1034-48. -   Chong et al., “PD1 blockade modulates chimeric antigen receptor     (CAR)-modified T cells: refueling the CAR,” Blood. 129(8), 1039-41,     2017, published on-line Dec. 28, 2016 -   Chowdhury et al., “Combination therapy strategies for improving PD1     blockade efficacy: a new era in cancer immunotherapy,” J. Int. Med.     doi: 10.1111/joim.12708, Epub ahead of print, Oct. 26, 2017 -   Creative Biolabs User Manual, “TriCo-20TM Phage Display 20-mer     Random Peptide Library,” 14 pages, Aug. 4, 2009 -   Dahlman et al., “In vivo endothelial siRNA delivery using polymeric     nanoparticles with low molecular weight,” Nat. Nanotechnol. 9,     648-55, 2014 -   Dempke et al., “Second- and third-generation drugs for     immuno-oncology treatment—The more the better?” Eur. J. Cancer 74,     55-72, March 2017 -   Desigaux et al., “Self-assembled lamellar complexes of siRNA with     lipidic aminoglycoside derivatives promote efficient siRNA delivery     and interference,” Proc. Nat'l. Acad. Sci. (USA) 104, 16534-39, 2007 -   Differding, “AUNP-12—A Novel Peptide Therapeutic Targeting PD1     Immune Checkpoint Pathway for Cancer Immunotherapy—Structure     Activity Relationships & Peptide/Peptidomimetic Analogs,” available     at     differding.com/data/AUNP_12_A_novel_peptide_therapeutic_targeting_PD_1_immune     checkpoint_pathway for cancer immunotherapy.pdf, Feb. 26, 2014 -   Dong et al., “Lipopeptide nanoparticles for potent and selective     siRNA delivery in rodents and nonhuman primates,” Proc. Nat'l. Acad.     Sci. (USA) 111, 3955-60, 2014 -   Dosta et al., “Surface charge tunability as a powerful strategy to     control electrostatic interaction for high efficiency silencing,     using tailored oligopeptide-modified poly(beta-amino ester)s     (PBAEs),” Acta Biomater. 20, 82-93, 2015 -   Duraiswamy et al., “Dual Blockade of PD1 and CTLA-4 Combined with     Tumor Vaccine Effectively Restores T-Cell Rejection Function in     Tumors,” Cancer Res 73, 3591-603, 2013 -   Fenton et al., “Bioinspired alkenyl amino alcohol ionizable lipid     materials for highly potent in vivo mRNA delivery,” Adv. Mater. 28,     2939-43, 2016 -   Feridooni et al., “Noninvasive Strategies for Systemic Delivery of     Therapeutic Proteins—Prospects and Challenges,” Chapter 8 of Sezer,     ed., Smart Drug Delivery System, available at     http://www.intechopen.com/books/smart-drug-delivery-system, Feb. 10,     2016 -   Freeman et al., “Phase I/II trial of intravenous NDV-HUJ oncolytic     virus in recurrent glioblastoma multiforme,” Mol. Ther. 13, 221-28,     2006 -   Gao et al., “VISTA is an inhibitory immune checkpoint that is     increased after ipilimumab therapy in patients with prostate     cancer,” Nature Med. 23, 551-55, 2017 -   Geevarghese et al., “Phase I/II Study of Oncolytic Herpes Simplex     Virus NV1020 in Patients with Extensively Pretreated Refractory     Colorectal Cancer Metastatic to the Liver,” Hum. Gene Ther. 21,     1119-28, 2010 -   Guo et al., “Systemic delivery of therapeutic small interfering RNA     using a pH-triggered amphiphilic poly-L-lysinenanocarrier to     suppress prostate cancer growth in mice,” Eur. J. Pharm. Sci. 45,     521-32, 2012 -   Harvey et al., “Efficacy of anti-ICOS agonist monoclonal antibodies     in preclinical tumor models provides a rationale for clinical     development as cancer immunotherapeutics,” Journal for ImmunoTherapy     of Cancer 3(Suppl 2), 09, 2015 -   He et al., “Lymphocyte-activation gene-3, an important immune     checkpoint in cancer,” Cancer Sci. 107, 1193-97, 2016 -   Howard et al., “RNA interference in vitro and in vivo using a novel     chitosan/siRNA nanoparticle system,” Mol. Ther. 14, 476-84, 2006 -   Huseni et al., “Anti-tumor efficacy and biomarker evaluation of     agonistic anti-OX40 antibodies in preclinical models,” Journal for     ImmunoTherapy of Cancer 2(Suppl 3), P105, 2014 -   Infante et al., “A phase Ib dose escalation study of the OX40     agonist MOXR0916 and the PD-L1 inhibitor atezolizumab in patients     with advanced solid tumors,” J Clin Oncol. 34(suppl; abstr 101),     2016 -   John et al., “Blockade of PD1 immunosuppression boosts CAR T-cell     therapy,” Oncolmmunology 2, e26286, 3 pages, 2013 -   Johnson et al., “A Cancer Research UK phase I study evaluating     safety, tolerability, and biological effects of chimeric anti-CD40     monoclonal antibody (MAb), Chi Lob 7/4,” J Clin Oncol. 28, 2507,     2010. -   Johnson et al., “Clinical and Biological Effects of an Agonist     Anti-CD40 Antibody: A Cancer Research UK Phase I Study,” Clin Cancer     Res 21, 1321-28, 2015 -   Judge & MacLachlan, “Overcoming the innate immune response to small     interfering RNA,” Hum Gene Ther. 2008; 19:111-24. -   Kaczmarek et al., “Advances in the delivery of RNA therapeutics:     from concept to clinical reality,” Genome Medicine 2017; 9:60, 16     pages -   Kanasty et al., “Delivery materials for siRNA therapeutics,” Nat.     Mater. 12, 967-77, 2013 -   Kauffman et al., “Optimization of lipid nanoparticle formulations     for mRNA delivery in vivo with fractional factorial and definitive     screening designs,” Nano Lett. 15, 7300-06, 2015 -   Kauffman et al., “Efficacy and immunogenicity of unmodified and     pseudouridine-modified mRNA delivered systemically with lipid     nanoparticles in vivo,” Biomaterials. 2016; 109:78-87. -   Kaufmann et al., “Chemovirotherapy of Malignant Melanoma with a     Targeted and Armed Oncolytic Measles Virus,” J. Invest. Dermatol.     133, 1034-42, 2013 -   Kavikansky & Pavlick, “Beyond Checkpoint Inhibitors: The Next     Generation of Immunotherapy in Oncology,” Amer. J. Hematol. Oncol.     13, 9-20, 2017 -   Khubchandani et al., “Dacetuzumab, a humanized mAb against CD40 for     the treatment of hematological malignancies,” Curr Opin Investig     Drugs 10, 579-87, 2009. -   Khuri et al., “A controlled trial of intratumoral ONYX-015, a     selectively-replicating adenovirus, in combination with cisplatin     and 5-fluorouracil in patients with recurrent head and neck cancer,”     Nat. Med. 6, 879-85, 2000 -   Kontermann, “Half-life extended biotherapeutics,” Expert Opin. Biol.     Ther. 16, 903-15, 2016. -   Kozielski et al., “A bioreducible linear poly(β-amino ester) for     siRNA delivery,” Chem. Commun. (Camb). 49, 5319-21, 2013 -   Lawler et al., “Oncolytic Viruses in Cancer Treatment,” JAMA Oncol.     3, 841-49, 2017 (published on-line Jul. 21, 2016) -   Le Mercier et al., “VISTA Regulates the Development of Protective     Antitumor Immunity,” Cancer Res 2014; 74:1933-1944 -   Leone et al., “A2aR antagonists: Next generation checkpoint blockade     for cancer immunotherapy,” Computational and Structural     Biotechnology Journal 13, 265-72, 2015 -   Leus et al., “VCAM-1 specific PEGylated SAINT-based lipoplexes     deliver siRNA to activated endothelium in vivo but do not attenuate     target gene expression,” Int. J. Pharm. 469, 121-31, 2014 -   Li et al., “Discovery of peptide inhibitors targeting human     programmed death 1 (PD1) receptor,” Oncotarget 7, 64967-76, Aug. 12,     2016 -   Li et al., “Effects of chemically modified messenger RNA on protein     expression,” Bioconjug Chem. 2016; 27:849-53. -   Liang, “Oncorine, the World First Oncolytic Virus Medicine and its     Update in China,” Curr. Cancer Drug Targets 18, 171-76, 2018 -   Linch et al., “OX40 agonists and combination immunotherapy: putting     the pedal to the metal,” Frontiers in Oncology 5, 14 pages, 2015 -   Liu et al., “Immune-checkpoint proteins VISTA and PD1 nonredundantly     regulate murine T-cell responses,” Proc. Nat'l. Acad. Sci. USA 112,     6682-87, 2015 -   Lorence et al., “Phase 1 clinical experience using intravenous     administration of PV701, an oncolytic Newcastle disease virus,”     Curr. Cancer Drug Targets 7, 157-67, 2007 -   Lorenz et al., “Steroid and lipid conjugates of siRNAs to enhance     cellular uptake and gene silencing in liver cells,” Bioorganic Med.     Chem. Lett. 14, 4975-77, 2004 -   Love et al., “Lipid-like materials for low-dose, in vivo gene     silencing,” Proc. Nat'l. Acad. Sci. (USA) 107, 1864-69, 2010 -   Lu et al., “Replicating retroviral vectors for oncolytic virotherapy     of experimental hepatocellular carcinoma,” Oncol. Rep. 28, 21-26,     2012 -   Lundstrom, “Oncolytic Alphaviruses in Cancer Immunotherapy,”     Vaccines 5, pages 1-17, 2017 -   Lynn & Langer, “Degradable poly(β-amino esters): synthesis,     characterization, and self-assembly with plasmid DNA,” J. Am. Chem.     Soc. 122, 10761-18, 2000 -   Magiera-Mularz et al., “Bioactive macrocyclic inhibitors of the     PD1/PD-L1 immune checkpoint,” Angewandte Chemie Int. Ed.     10.1002/anie.201707707, e-published Sep. 26, 2017 -   Maute et al., “Engineering high-affinity PD1 variants for optimized     immunotherapy and immuno-PET imaging,” Proc. Natl. Acad. Sci. USA,     E6506-E6514, published online Nov. 10, 2015 -   McDonald et al., “A measles virus vaccine strain derivative as a     novel oncolytic agent against breast cancer,” Breast Cancer Treat.     99, 177-84, 2006 -   McOmie, Protective Groups in Organic Chemistry, Plenum Press, New     York, N.Y., 1973 -   Mediavilla-Varela et al., “A Novel Antagonist of the Immune     Checkpoint Protein Adenosine A2a Receptor Restores     Tumor-Infiltrating Lymphocyte Activity in the Context of the Tumor     Microenvironment,” Neoplasia 19, 530-36, 2017 -   Mellemgaard et al., “Combination immunotherapy with IDO vaccine and     PD1 inhibitors in advances HSCLC,” DOI: 10.1200/JCO.2017.35.15     suppl.TPS2610 Journal of Clinical Oncology 35, no.     15_suppl—published online before print, 2017 -   Merrifield, “Solid phase peptide synthesis I: Synthesis of a     tetrapeptide,” J. Am. Chem. Soc. 85:2149-54, 1963 -   Messenheimer et al., “Timing of PD1 Blockade Is Critical to     Effective Combination Immunotherapy with Anti-OX40,” Clin. Cancer     Res. 23, DOI: 10.1158/1078-0432.CCR-16-2677 Published October 2017 -   Michaelson et al., “Preclinical evaluation of JTX-2011, an anti-ICOS     agonist antibody,”, Abstract 573, Proceedings: AACR 107th Annual     Meeting 2016; Apr. 16-20, 2016; New Orleans, La. -   Morrissey et al., “Immunotherapy and Novel Combinations in Oncology:     Current Landscape, Challenges, and Opportunities,” Clinical and     Translational Science 9, 89-104, 2016 -   Morrissey et al., “Potent and persistent in vivo anti-HBV activity     of chemically modified siRNAs,” Nat. Biotechnol. 23, 1002-07, 2005 -   Moschos et al., “Lung delivery studies using siRNA conjugated to     TAT(48-60) and penetratin reveal peptide induced reduction in gene     expression and induction of innate immunity,” Bioconjug. Chem. 18,     1450-59, 2007 -   Nair et al., “Multivalent N-acetylgalactosamine-conjugated siRNA     localizes in hepatocytes and elicits robust RNAi-mediated gene     silencing,” J. Am. Chem. Soc. 136, 16958-61, 2014 -   Neurath et al., eds., The Proteins, Vol. II, 3d ed., pp. 105-237,     Academic Press, New York, N.Y. (1976) -   Nishina et al., “Efficient in vivo delivery of siRNA to the liver by     conjugation of alphatocopherol.,” Mol. Ther. 16, 734-40, 2008 -   Ott et al., “Combination immunotherapy: a road map,” J.     ImmunoTherapy of Cancer 5, 16, 2017 -   Pack et al., “Design and development of polymers for gene delivery,”     Nat. Rev. Drug discov. 4, 581-93, 2005 -   Patel et al., “Recent Advances in Protein and Peptide Drug Delivery:     A Special Emphasis on Polymeric Nanoparticles,” Protein. Pept. Lett.     21, 1102-20, 2014 -   Penchala et al., “A biomimetic approach for enhancing the in vivo     half-life of peptides,” Nat. Chem. Biol. 11, 793-98, 2015 -   Phuangsab et al., “Newcastle disease virus therapy of human tumor     xenografts: antitumor effects of local or systemic administration,”     Cancer Lett. 172, 27-36, 2001 -   Prakash et al., “Positional effect of chemical modifications on     short interference RNA activity in mammalian cells,” J Med Chem.     2005; 48:4247-53 -   Pratt & MacRae, “The RNA-induced silencing complex: a versatile     gene-silencing machine,” J Biol Chem. 2009; 284:17897-901 -   Rehman et al., “Mechanism of polyplex- and lipoplexmediated delivery     of nucleic acids: real-time visualization of transient membrane     destabilization without endosomal lysis,” ACS Nano. 7, 3767-77, 2013 -   Rivera et al., “Hair Repigmentation During Immunotherapy Treatment     With an Anti-Programmed Cell Death 1 and Anti-Programmed Cell Death     Ligand 1 Agent for Lung Cancer,” JAMA Dermatol. 2017 Jul. 12. doi:     10.1001/jamadermatol.2017.2106, Jul. 12, 2017 -   Rudin et al., “Phase I clinical study of Seneca Valley Virus     (SVV-001), a replication-competent picornavirus, in advanced solid     tumors with neuroendocrine features,” Clin. Cancer Res. 17, 888-95,     2011 -   Sahin et al., “mRNA-based therapeutics—developing a new class of     drugs,” Nat Rev Drug Discov. 2014; 13:759-80 -   Sakuishi et al., “Targeting Tim-3 and PD1 pathways to reverse T cell     exhaustion and restore anti-tumor immunity,” J. Exp. Med. 20,     2187-94, 2010 -   Schaer et al., “Modulation of GITR for cancer immunotherapy,” Curr     Opin Immunol. 24, 217-24, 2012 -   Schroeder et al., “Lipid-based nanotherapeutics for siRNA     delivery,” J. Int. Med. 267, 9-21, 2010 -   Sharma & Allison, “Immune Checkpoint Targeting in Cancer Therapy:     Toward Combination Strategies with Curative Potential,” Cell 161,     205-14, 2015 -   Shindo et al., “Combination Immunotherapy with 4-1BB Activation and     PD1 Blockade Enhances Antitumor Efficacy in a Mouse Model of     Subcutaneous Tumor,” Anticancer Res. 35, 129-36, 2015 -   Shrimali et al., “Concurrent PD1 Blockade Negates the Effects of     OX40 Agonist Antibody in Combination Immunotherapy through Inducing     T-cell Apoptosis,” Cancer Immunol Res 5(9), pages OF1-12, Aug. 28,     2017 -   Skalniak et al., “Small-molecule inhibitors of PD1/PD-L1 immune     checkpoint alleviate the PD-L1-induced exhaustion of T-cells,”     Oncotarget, Advance Publications, Aug. 7, 2017, 15 pages -   Smith, “Pigmented skin lesions lightened during melanoma     immunotherapy,”     http://www.mdedge.com/edermatologynews/article/132598/melanoma/pigmented-skin-lesions-lightened-during-melanoma,     Mar. 2, 2017 -   Soutschek et al., “Therapeutic silencing of an endogenous gene by     systemic administration of modified siRNAs,” Nature. 2004;     432:173-78 -   Spodzieja et al., “Design of short peptides to block BTLA/HVEM     interactions for promoting anticancer T-cell responses,” PLoS ONE     12(6): e0179201, 17 pages, 2017 -   Stojdl et al., “Exploiting tumor-specific defects in the interferon     pathway with a previously unknown oncolytic virus,” Nat. Med. 6,     821-25, 2000 -   Stojdl et al., “VSV strains with defects in their ability to     shutdown innate immunity are potent systemic anti-cancer agents,”     Cancer Cell 4, 263-75, 2003 -   Stuart & Young, Solid Phase Peptide Synthesis, Pierce Chemical     Company, Rockford, Ill., 1984 -   Tigue et al., “MEDI1873, a potent, stabilized hexameric agonist of     human GITR with regulatory T-cell targeting potential,”     ONCOIMMUNOLOGY 6(3), e1280645 (14 pages), Feb. 3, 2017 -   Triebel et al., “LAG-3, a novel lymphocyte activation gene closely     related to CD4,” J. Exp. Med. 171, 1393-405, 1990 -   Tsutsumi et al., “Evaluation of polyamidoamine     dendrimer/alpha-cyclodextrin conjugate (generation 3, G3) as a novel     carrier for small interfering RNA (siRNA),” J. Control. Release 119,     349-59, 2007 -   Tuck, “Development of Small Molecule Checkpoint Inhibitors,” Immune     Checkpoint Inhibitors Symposium, 28 pages, Mar. 14-16, 2017 -   Tzeng et al., “Cystamine-terminated poly(beta-amino ester)s for     siRNA delivery to human mesenchymal stem cells and enhancement of     osteogenic differentiation,” Biomaterials 33, 8142-51, 2012 -   Tzeng et al., “PD1 blockage reverses immune dysfunction and     hepatitis B viral persistence in a mouse animal model,” PLoS One     7(6):e39179, 2012 -   Van Dessel et al., “Potent and tumor specific: arming bacteria with     therapeutic proteins,” Ther. Deliv. 6, 385-99, 2015 -   Vonderheide and Glennie, “Agonistic CD40 antibodies and cancer     therapy,” Clin. Cancer Res. 19, 1035-43, 2013 -   Vonderheide et al., “Clinical activity and immune modulation in     cancer patients treated with CP-870,893, a novel CD40 agonist     monoclonal antibody,” J Clin Oncol. 25, 876-83, 2007 -   Wang et al., “Anaplastic lymphoma kinase (ALK) inhibitors: a review     of design and discovery,” Med. Chem. Commun. 5, 1266-79, 2014 -   Wang et al., “VISTA, a novel mouse Ig superfamily ligand that     negatively regulates T cell responses,” J. Exp. Med. 208, 577-92,     2011 -   Wittrup & Lieberman, “Knocking down disease: a progress report on     siRNA therapeutics,” Nat Rev Genet. 2015; 16:543-52 -   Won et al., “Missing pieces in understanding the intracellular     trafficking of polycation/DNA complexes,” J. Control. Release 139,     88-93, 2009 -   Xia et al., “Antibody-mediated targeting of siRNA via the human     insulin receptor using avidin—biotin technology.,” Mol. Pharm. 6,     747-51, 2009 -   Yang et al., “Oral vaccination with salmonella simultaneously     expressing Yersinia pestis F1 and V antigens protects against     bubonic and pneumonic plague,” J Immunol. 178, 1059-67, 2007 -   Ye et al., “T-cell exhaustion in chronic hepatitis B infection:     current knowledge and clinical significance,” Cell Death Dis. 19,     e1694, 2015 -   Young et al., “Co-inhibition of CD73 and A2AR Adenosine Signaling     Improves Anti-tumor Immune Responses,” Cancer Cell 30, 391-403, 2016 -   Yu et al., “Disposition and pharmacology of a GalNAc3-conjugated ASO     targeting human lipoprotein(a) in mice,” Mol. Ther. Nucleic Acids 5,     e317, 2016 -   Zarganes-Tzitzikas et al., “Inhibitors of programmed cell death 1     (PD1): a patent review,” Expert Opinion on Therapeutic Patents 26,     973-77, published on-line Jul. 6, 2016 -   Zhan et al., “From monoclonal antibodies to small molecules: the     development of inhibitors targeting the PD1/PD-L1 pathway,” Drug     Discovery Today 21, 1027-36, June 2016 -   Zorzi et al., “Acylated heptapeptide binds albumin with high     affinity and application as tag furnishes long-acting peptides,”     Nature Communications 8, 16092, 2017 

1. A peptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3.
 2. An expression construct encoding a peptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, and SEQ ID NO:4.
 3. The expression construct of claim 2, which is present in a CAR-T cell or an oncolytic virus.
 4. A pharmaceutical composition comprising: (a) an active agent selected from the group consisting of: (i) a peptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, and SEQ ID NO:4; (ii) a nucleic acid encoding the peptide; (iii) a CAR-T cell expressing the peptide; (iv) an oncolytic virus expressing the peptide; and (b) a pharmaceutically acceptable carrier.
 5. The pharmaceutical composition of claim 4, wherein the active agent is the nucleic acid, wherein the nucleic acid is selected from the group consisting of DNA, cDNA, PNA, and RNA.
 6. The pharmaceutical composition of claim 5, wherein the nucleic acid is RNA.
 7. The pharmaceutical composition of claim 6, wherein the RNA comprises a modification selected from the group consisting of (i) modification of a ribose sugar, (ii) modification of a phosphate linkage, and (iii) modification of a base.
 8. The pharmaceutical composition of claim 7, wherein the modification is selected from the group consisting of a ribo-difluorotoluyl nucleotide, a 4′-thio modified RNA, a boranophosphate linkage, a phosphorothioate linkage, a 2′-O-methyl (2′-OMe) sugar substitution, a 2′-fluoro (2′-F), a 2′-O-methoxyethyl (2′-MOE) sugar substitution, a locked nucleic acid (LNA), and an L-RNA.
 9. The pharmaceutical composition of claim 4, wherein the active agent is the peptide, wherein the peptide is provided with a peptide carrier system selected from the group consisting of a microparticle, a polymeric nanoparticle, a liposome, a solid lipid nanoparticle, a hydrophilic mucoadhesive polymer, a thiolated polymer, a polymer matrix, a nanoemulsion, and a hydrogel.
 10. A method of inhibiting the progression of a hyperproliferative disorder, treating an infectious disease, enhancing a response to vaccination, treating sepsis, promoting hair re-pigmentation, or promoting lightening of a pigmented skin lesion, comprising administering to an individual in need thereof an effective amount of the pharmaceutical composition of claim
 4. 11. The method of claim 10, wherein the pharmaceutical composition is administered to inhibit progression of the hyperproliferative disorder.
 12. The method of claim 11, wherein the hyperproliferative disorder is a cancer.
 13. The method of claim 12, wherein the cancer is a pancreatic cancer or a colon carcinoma.
 14. The method of claim 10, further comprising administering a second therapy to the patient.
 15. The method of claim 14, wherein the second therapy is selected from the group consisting of: (i) a cancer vaccine; (ii) a chimeric antigen receptor (CAR) T cell therapy; (iii) a therapy that comprises reducing or blocking activity of a molecule selected from the group consisting of PD1, PD-L1, lymphocyte-activation gene-3 (LAG-3), cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4), V-domain Immunoglobulin Suppressor of T cell Activation (VISTA), T-cell Immunoglobulin domain and Mucin domain 3 (TIM-3), a killer immunoglobulin-like receptor (KIR), indoleamine (2,3)-dioxygenase (IDO), B and T Lymphocyte Attenuator (BTLA), A2A adenosine receptor (A2AR); (iv) a cytokine; (v) an agonist of a molecule selected from the group consisting of CD40, OX40, glucocorticoid-induced tumor necrosis factor-related protein (GITR), and Inducible T-cell COStimulator (ICOS); (vi) an oncolytic virus; and (vii) a therapeutic agent selected from the group consisting of a 4-1BB agonist, a 4-1BB antagonist, an inhibitor of anaplastic lymphoma kinase (ALK), an inhibitor of histone deacetylase (HDAC), and an inhibitor of VEGFR.
 16. The method of claim 10, wherein the pharmaceutical composition is administered to treat an infectious disease or as a vaccine adjuvant to a vaccine against the infectious disease.
 17. The method of claim 16, wherein the infectious disease is malaria or hepatitis B.
 18. The method of claim 10, wherein the at least one peptide is administered to treat sepsis.
 19. The method of claim 10, wherein the at least one peptide is administered to promote hair re-pigmentation or to promote lightening of a pigmented skin lesion. 